RNA-Seq of sorted Lgr5+ cells with and without infection with Helicobacter pylori
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE129107 | GEO | 2019/04/02
REPOSITORIES: GEO
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Project description:R-spondin (Rspo) signaling is crucial for stem cell renewal and tissue homeostasis in the gastrointestinal tract. In the stomach, Rspo is secreted from myofibroblasts and controls epithelial gland regeneration by inducing proliferation of Wnt-responsive Axin2+ cells. Infection with H. pylori results in increased expression of stromal Rspo, leading to an expansion of Axin2+ isthmus stem cells and gland hyperplasia. Lgr5+ stem cells are exposed to Rspo3 but the effects of this are not well understood. Here we demonstrate that in addition to its effect as a mitogen, endogenous Rspo3 regulates the gene expression of Lgr5+ cells in the gland base in both antrum and corpus. Rspo3 inhibits proliferation and induces differentiation within the Lgr5+ compartment towards a secretory phenotype. Strikingly, the Rspo3-Lgr5 axis turns out to be required for epithelial antimicrobial defense. Infection with H. pylori induces a strong antimicrobial response, with Lgr5+ cells expressing antimicrobial compounds that are secreted into the lumen in an Rspo3-dependent manner. Depletion of Lgr5+ cells or knockout of Rspo3 in myofibroblasts leads to dramatic hyper-colonization of gastric glands, including the stem cell compartment, whereas systemic application of recombinant Rspo is sufficient to clear H. pylori from the glands. We provide an intriguing, unexpected feature of Lgr5+ cells, exhibiting an indivisible connection between stem cell signaling and antimicrobial self-protection.
2019-01-01 | GSE108729 | GEO
Project description:R-spondin (Rspo) signaling is crucial for stem cell renewal and tissue homeostasis in the gastrointestinal tract. In the stomach, Rspo is secreted from myofibroblasts and controls epithelial gland regeneration by inducing proliferation of Wnt-responsive Axin2+ cells in the isthmus of the gland. Infection with H. pylori results in increased expression of stromal Rspo, leading to an expansion of Axin2+ isthmus stem cells and gland hyperplasia. Lgr5+ cells in the gland base are exposed to Rspo3 but the effects of this are not well understood. Here we demonstrate that apart from its activity as a mitogen, endogenous Rspo3 regulates gene expression of Lgr5+ cells in the gastric gland base. Surprisingly, Rspo3 induces differentiation within the Lgr5+ compartment towards secretory deep mucous cells. Moreover, the Rspo3-Lgr5 axis turns out to be a stimulus of epithelial antimicrobial defense. Infection with H. pylori induces a strong antimicrobial response, with Lgr5+ cells expressing antimicrobial compounds that are secreted into the lumen in an Rspo3-dependent manner. Depletion of Lgr5+ cells or knockout of Rspo3 in myofibroblasts leads to hyper-colonization of gastric glands, including the stem cell compartment, whereas systemic application of recombinant Rspo clears H. pylori from the glands. We provide an intriguing, unexpected feature of the Rspo3-Lgr5 axis in the stomach, exhibiting antimicrobial self-protection of the gland to protect the stem cell compartment from invading pathogens.
2019-03-25 | GSE124930 | GEO
Project description:The gastrointestinal epithelial crypts are clonal units with a high cell turnover, driven by a small population of long-lived, Lgr5-expressing stem cells located in the crypt base. Despite this, depletion of Lgr5+ cells does not lead to severe pathology. Instead, other cell populations, such as secretory and enterocyte precursors are able to de-differentiate, replace Lgr5+ cells, and regenerate the crypt. However, the signals that regulate this epithelial plasticity are not well understood. Here we illuminate the hierarchical organization and regulation of stem cell plasticity in the colonic crypt. Using in vivo lineage tracing, we find that the classic Wnt target gene Axin2 is expressed in colonic Lgr5+ cells and adjacent Lgr5-negative cells that give rise to entire crypts upon Lgr5+ cell depletion. The identity of Lgr5+ cells is controlled by by R-spondin 3 (Rspo3) produced by myofibroblasts and the recovery of Lgr5+ cells upon depletion required functional Rspo3 signaling. In contrast, expression of Axin2 and the reserve stem cell function of Axin2+ cells is not dependent on Rspo3. Accordingly, upon knockout of Rspo3, Lgr5-negative Axin2+ cells are able to compensate the loss of the Lgr5+ cell compartment and maintain epithelial integrity. In contrast, Rspo3 is essential for maintaining epithelial integrity in the context of injury of the entire Axin2+ cell pool, as observed in the context of DSS colitis. We demonstrate that DSS induces a progressive loss of the Lgr5+ stem cells as well as Axin2+ Lgr5-negative reserve stem cell compartment. While Rspo3 deficient animals are unable to withstand this injury, in Rspo3 competent animals, crypt homeostasis and regeneration are fueled by the remaining Axin2-negative, differentiated cells. These cells can be reprogrammed by Rspo3 to proliferate and act as stem cells. Thus, we identity Rspo3 as a major regulator of the Lgr5 stem cell signature. While loss of this signature can be compensated by Rspo3-independent reserve stem cells under healthy conditions, it is essential for re-establishment of epithelial integrity upon crypt injury.
2019-06-01 | GSE115752 | GEO
Project description:Gastric epithelial stem cells are responsible for constant epithelial self-renewal, which is accelerated by infection with the gastric pathogen Helicobacter pylori. However, the mechanism that regulates stem cell turnover in the stomach remains unknown. Here we show that signaling by R-spondin 3 and Wnt hierarchically organizes the stem cell compartment in the antrum, producing two Wnt-responsive populations, which are either Lgr5+ve or Axin2 +ve. The positional identity of the Axin2+ve population relies on R-spondin 3 produced by stromal myofibroblasts. Increased availability of R-spondin induces hyperproliferation through specific expansion of Axin2+ve but not Lgr5+ve cells. Similarly, infection with H. pylori induces an increase in stromal R-spondin 3 expression, resulting in hyperplasia as well as shedding of bacteria that have entered the gland. This identifies a role for stromal cells in environmental sensing to orchestrate epithelial homeostasis via Wnt signaling.
2017-06-30 | GSE79494 | GEO
Project description:RSPO is a WNT pathway activator and functions as a potent regulator of stem cell growth in colon. RSPO family members were produced by several human tumors representing multiple tumor types including ovarian, pancreatic, colon, breast and lung cancer. Specific monoclonal antibody antagonists of RSPO family members were developed. In human patient-derived tumor xenograft models, anti-RSPO treatment markedly inhibited tumor growth either as single agents or in combination with chemotherapy. Furthermore, blockade of RSPO signaling reduced the tumorigenicity of cancer cells based on serial transplantation studies. In order to assess the impact of RSPO3 inhibition and gain insight in the anti-RSPO3 treatment mechanism of action, the global gene expression profiles of 4 human colorectal cancer patient derived models (PDX) were performed using Affymetrix microarray for the xenografts treated by the anti-RSPO3 antibody.
2016-03-03 | GSE73906 | GEO
Project description:The glandular stomach is comprised of two anatomically and functionally distinct epithelial tissues termed corpus and antrum. While these compartments continuously regenerate to ensure normal organ function, our understanding of the molecular and cellular determinants that maintain these tissues remains incomplete. Here, we established an unbiased account of all gastric epithelial cells of the mouse corpus and antrum using single cell RNA-sequencing, allowing us to compare these adjacent epithelia and their stem cell compartments. Our data reveals that epithelial cell types with equivalent functional roles in the corpus and antrum utilize similar transcriptional programs, and our results support the existence of two discernible stem cell populations within the gland base and isthmus regions of these tissues. To study isthmus stem cells, typically underrepresented in conventional, gland base-enriched organoid cultures, we developed a novel 2D monolayer culture system to propagate mouse and human gastric epithelia. These 2D cultures expand indefinitely via Lgr5- isthmus-like cells and maintain the ability to differentiate into diverse gastric cell types. While 2D corpus and antrum cultures were transcriptionally similar, they retained a molecular memory and differentiation bias that mirrored their tissue of origin. Furthermore, gastric 2D cultures could be readily and reversibly converted into conventional 3D organoids, highlighting the remarkable plasticity of undifferentiated stomach epithelial cells. Finally, we utilized the 2D culture system to show that rare primary epithelial cells with high levels of Sox2 expression, previously hypothesized to be gastric stem cells, most closely resembled enterochromaffin (EC) cells and Sox2 is both necessary and sufficient to generate EC cells in vivo. Together, our data (i) provide important insights into the basis of gastric epithelial regeneration and differentiation, (ii) establish a tractable 2D culture system to capture and manipulate primary gastric isthmus stem cells in vitro and (iii) uncover a role for the stem cell factor Sox2 during EC cell specification.
2022-11-10 | GSE217472 | GEO
Project description:Chief cells expressing Wnt/R-spondin target genes are located in the stomach corpus gland base. These cells can act as reserve stem cells and their transdifferentiation is a hallmark of spasmolytic polypeptide expressing metaplasia (SPEM). Mechanisms that control their differentiation, transdifferentiation and re-differenitiation are not fully understood. We investigated the function of R-spondin 3, a Wnt signaling activator and Lgr5 ligand, by using conditional mouse models that enable manipulation of R-spondin 3 gene expression in gastric myofibroblasts.The effect and the physiological function of R-spondin 3 was analyzed in the corpus and colon with R-spondin 3 gene knock-out mice in Myh11+ myofibroblasts (Myh11-CreERT2; Rspo3fl/fl (RSPO3 KO) or R-spondin 3 gene knock-in overexpressiong myofibroblasts ( Myh11-CreERT2/Rosa26Sor6(CAG–Rspo3) ( RSPO3 KI)).
2022-01-01 | GSE145087 | GEO
Project description:The glandular stomach is comprised of two regenerative compartments termed corpus and antrum, yet our understanding of the cell populations that maintain these epithelia is incomplete. Here, we show that cell types with equivalent functional roles in the corpus and antrum share similar transcriptional states including the immature isthmus population. To study isthmus cells, we developed a monolayer (2D) culture that is continually maintained via Lgr5-negative cells and produces diverse gastric cell types. Importantly, 2D cultures can be interconverted into conventional 3D organoids, recapitulating the plasticity of gastric epithelial cells in vivo. Finally, we utilized the 2D culture system to show that rare Sox2-expressing cells resemble enterochromaffin (EC) cells and Sox2 is both necessary and sufficient to generate EC cells. Together, our data provide important insights into the basis of gastric homeostasis, establish a tractable culture system to capture isthmus cells and uncover a role for Sox2 in EC cells.
2022-11-10 | GSE217471 | GEO
Project description:The glandular stomach is comprised of two regenerative compartments termed corpus and antrum, yet our understanding of the cell populations that maintain these epithelia is incomplete. Here, we show that cell types with equivalent functional roles in the corpus and antrum share similar transcriptional states including the immature isthmus population. To study isthmus cells, we developed a monolayer (2D) culture that is continually maintained via Lgr5-negative cells and produces diverse gastric cell types. Importantly, 2D cultures can be interconverted into conventional 3D organoids, recapitulating the plasticity of gastric epithelial cells in vivo. Finally, we utilized the 2D culture system to show that rare Sox2-expressing cells resemble enterochromaffin (EC) cells and Sox2 is both necessary and sufficient to generate EC cells. Together, our data provide important insights into the basis of gastric homeostasis, establish a tractable culture system to capture isthmus cells and uncover a role for Sox2 in EC cells.
2022-11-10 | GSE217470 | GEO
Project description:Biliary epithelial cells (BECs) form bile ducts in the liver, and are facultative liver stem cells that establish a ductular reaction (DR) to support liver regeneration following injury. Liver damage induces periportal LGR5+ putative liver stem cells which can form BEC-like organoids, suggesting RSPO-LGR4/5-mediated WNT/β-catenin activity is important for a DR. We addressed the roles for this and other signaling pathways in a DR by performing a focused CRISPR-based loss-of-function screen in BEC-like organoids, followed by in vivo validation and single-cell RNA sequencing. We found BECs lack and do not require LGR4/5-mediated WNT/β-Catenin signaling during DR, while YAP and mTORC1 signaling are required for this process. Upregulation of LGR5/AXIN2 is required in hepatocytes to enable their regenerative capacity in response to injury. Together, these data highlight heterogeneity within the BEC pool and delineate signaling pathways involved.
2018-08-27 | GSE111269 | GEO